JP2000088805A - Column connecting joint, capillary column and capillary electric chromatography device using same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To rapidly and simply connect a capillary column by constituting a column connecting joint by a body having a hole of a small diameter and a frit provided in an internal through hole; screws used on both ends; and a ferrule. SOLUTION: A body 13 of a column connecting joint is selected from various kinds of glasses, a resin or the like and has a hole 14 of a small diameter for connecting a capillary tube 24 for an electrode cell at a sample pouring side at one end. The body 13 has a hole 15 of a small diameter for connecting a capillary column 25 filling a filler at the other end and both holes 14, 15 are communicated with each other to form an internal through hole 16. The capillary column 25 is connected to the internal through hole 16 and a projecting part 27 for fixing a frit 19 retaining the filler 26 is provided on the through hole 16. Screws 20, 21 engaged with engaging screws 17, 18 threaded on both inner periphery of the body 13 have internal through holes. One hole is the one for inserting the capillary tube 24 for an electrode cell at a sample-pouring side and the other hole is the one for inserting the capillary column 25 respectively. Ferrules 22, 23 are interposed between these screws 20, 21 and the body 13.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a capillary electrophoresis apparatus (hereinafter abbreviated as "CE apparatus") used for capillary electrophoresis (hereinafter abbreviated as "CE"), and more particularly, to an electroosmotic flow (hereinafter abbreviated as "CE apparatus"). Capillary electrochromatography (hereinafter abbreviated as “EOF”) that performs separation analysis based on the interaction between a sample and a stationary phase and the difference in electric mobility in a capillary column packed with a packing material using a driving force of a mobile phase. The present invention relates to a column connection joint suitably used for “CEC”), a capillary column provided with the column connection joint,
Further, the present invention relates to a CEC device provided with such a column connection joint and a capillary column.

[0002]

2. Description of the Related Art CE is a separation / analysis method having excellent characteristics such as high absolute sensitivity, an extremely small amount of sample and a very small amount of reagent required for analysis. Generally CE
Is the difference in electric mobility between sample ions. The separation field is 0.1 mm or less in inner diameter and several tens cm to 1 m in length.
A degree of fused silica tube (hereinafter abbreviated as “capillary tube”) is used. By using a capillary tube, a high voltage of up to about 30 kV can be applied to generate a high potential gradient, so that rapid analysis is possible. Further, since the flow of the liquid sent in the CE becomes a plug flow due to the influence of EOF, a high theoretical stage number can be expected.

[0003] On the other hand, CEC uses basically the same device as CE, and a stationary phase used for high-performance liquid chromatography (hereinafter abbreviated as "HPLC") is filled in a capillary tube or the like for a measurement column. It is a new separation and analysis means that uses a capillary column and EOF instead of an HPLC pump for sending the mobile phase. In the measurement using such CEC, charged sample components are separated based on the interaction with the stationary phase in the capillary column and the difference in electric mobility, so that the separation selectivity is higher than that of HPLC. Is expected to improve. In addition, for the CEC of a chemical species that has no charge,
Separation will be achieved by the same mechanism as HPLC. The EOF generated by the electric field is a plug flow in the capillary column as in the case of CE, which is in contrast to the laminar flow in the case of pumping. Even if used, an increase in pressure due to liquid transfer does not matter. Therefore, CEC can be considered as an analytical method that combines high theoretical plate number of CE and high selectivity of HPLC.

[0004] In the capillary column used in the CEC, it is necessary to install a frit for holding the packing material in the column. Until now, 1) packing glass wool (J. Chromatgr., Vol. 213, p. 25) , 19
In 1981, J. Chromatgr. , 242, 33
1, 1982), 2) Insert a porous porous polymer (Anal. Chem., 56, 299).
0, 1984) 3) Glass beads packed and baked by arc discharge (J. Chroma)
tgr. 593, 313, 1992).
4) A mixture of silicate silica and formamide is inserted into a capillary column and polymerized therein (HRC & C)
C, 10, 446, 1987) has been used as a method for producing a frit. As described above, the production of the micro frit is performed by using a capillary column for CEC, or by using an HPLC using a capillary column.
(Hereinafter abbreviated as “CHPLC”). However, it is difficult to precisely control the pore diameter and length of the frit in any of the above-mentioned preparation methods, and it is necessary to prepare a frit for each capillary column. When filling with 1)
There is a problem that the filling operation becomes complicated because operations such as preparation of a column intermediate frit, 2) filling of a filler, and 3) preparation of a column end frit are required.

Further, a capillary column packed with a packing material used in the CEC has a structure in which a packing column structure in which the packing material is held by a frit and a cell structure for detecting a sample component by a detector are integrated. Therefore, after the filling operation into the capillary tube is completed, it is necessary to prepare a cell for detection for each capillary column, and the mounting and replacement of the capillary column with the CE device becomes complicated. was there.

Further, a capillary column packed with a filler used in CEC has a stationary phase for HPLC, and thus can be applied to CHPLC, but the capillary column structure does not have a connection structure to other equipment. For C
When connecting to an HPLC device, a new connecting part is required. Therefore, the current capillary column structure is limited to a simple CEC capillary column, and there is a problem that the field of application of the capillary column is narrow.

As described above, in the conventional capillary column, 1) it is necessary to prepare a frit and a detection cell for each capillary column; 2) there is no efficient packing method of the capillary column including the preparation of the frit;
3) lack of quickness and simplicity in measurement operation, etc.
4) There were problems such as the narrow application field of the capillary column.

[0008]

SUMMARY OF THE INVENTION An object of the present invention is to solve the above problems and the like.
To provide a column connection joint that can quickly and easily connect the capillary columns used in the test, has excellent insulation properties, and can minimize the diffusion of the sample during measurement and increase the resolution. It is in. In addition, using this column connection joint, the filling operation including the production of a frit is easy, and various fillers can be used in combination. Another object of the present invention is to provide a CEC device provided with a column connection joint and a capillary column.

[0009]

Means for Solving the Problems The present inventors have conducted intensive studies to solve the above-mentioned problems, and as a result, the conventional capillary column structure was replaced with a capillary tube for an electrode tank on the sample injection side,
By dividing into a capillary column filled with a filler and a capillary tube for the electrode tank on the detector side and connecting them with a column connection joint having a frit inside, 1) the production of frit and detection cell for each capillary column can be omitted. It has been found that it can be simplified, 2) exchange of capillary columns and connection between various capillary columns are easy, and 3) it can be applied as an analytical column even in CHPLC. Further, the present inventors have found that if a column in which the column connection joint is connected to a capillary column in advance is prepared, an appropriate analytical system can be set by exchanging the column or the like according to measurement conditions, thereby completing the present invention. Reached.

That is, the present invention relates to a capillary electrophoresis apparatus provided with means for separating a sample introduced from an end of a capillary column by applying a high voltage to both ends of the capillary column and detecting the separated sample components. , A structure in which capillary tubes for the sample injection side electrode tank and the detector side electrode tank are connected to both sides of the capillary column filled with the filler via connection joints each having a frit therein, and the connection joints. However, one has a small-diameter hole for connecting a capillary column filled with a filler, and the other has a small-diameter hole for connecting a capillary tube. A through-hole is formed, and a frit for holding the filler inside the internal through-hole is on the capillary column side. A connecting joint main body in which screw screws are engraved on inner peripheral portions of both ends, respectively, a screw having a through hole to be screwed into both ends of the connecting joint main body, and the connecting joint main body And a ferrule interposed between the screw and the screw, and the material of the connection joint has electrical insulation.

Hereinafter, the present invention will be described in detail.

First, the column connection joint of the present invention will be described. The column connection joint comprises a main body, a frit, a screw, and a ferrule. First, the main body will be described.

<Joint body for column connection> As the material of the body, 1) it must have electrical insulation properties, 2) it must not be corroded by contact with an electrolytic solution, and 3) it can sufficiently withstand Joule heat generated in the column. 4) There is no particular limitation as long as it can be molded as a connection joint.
For example, various glasses such as quartz and hard glass, polyethylene, polypropylene, polystyrene, polyvinyl chloride,
Various resins such as polyethylene terephthalate, polytetrafluoroethylene, polyimide, polyamide, polyetheretherketone, phenolic resin, and fluororesin, and various ceramics such as alumina, zirconia, titania, and silicon nitride can be appropriately selected and used. Further, depending on the structure and shape of the column connection joint of the present invention, not only one kind alone, but two or more kinds may be used in combination. Of these, resins are preferably used because of ease of molding, and polyetheretherketone is more preferably used.

As for the shape of the main body, a small-diameter hole for connecting a capillary tube for the sample-injection-side electrode tank or a capillary tube for the detector-side electrode tank is provided on one side as an internal through-hole. The diameter of the small-diameter hole is preferably slightly larger than the outer diameter of the capillary tube used in order to make the capillary tube adhere to the main body,
More preferably, the diameter can be appropriately selected from the range of about 0.25 to 0.4 mm. Further, the length of the small diameter hole is not particularly limited as long as it does not hinder the connection work, but can be appropriately selected from a range of about 0.5 to 1.0 cm.

The other end of the main body has a small-diameter hole for connecting a capillary column filled with a filler, and the small-diameter hole and the small-diameter hole are connected to each other to form an internal through-hole. ing. The diameter of the small-diameter hole is preferably slightly larger than the outer diameter of the capillary column to be used in order to bring the capillary column into close contact with the main body, and more preferably can be appropriately selected from the range of about 0.25 to 0.4 mm in diameter. . The length of the small diameter hole is not particularly limited as long as it does not hinder the connection work.
It can be appropriately selected from the range of 1.0 to 1.0 cm.

The length of the main body is not particularly limited and may be appropriately selected according to the purpose, but it may be desirable to select the length from a range of about 1 to 3 cm from the viewpoint of ease of connection work and the like. .

Further, a convex portion for fixing a frit used for connecting a capillary column can be provided in the internal through hole of the main body, and any shape may be used as long as the frit can be fixed. It is preferable that the inner diameter of the electrolyte be larger than the inner diameter of the electrolyte, because the electrolyte, which becomes a mobile phase at the time of measurement, can move quickly. The thickness of the projection depends on the material of the main body, but may be any thickness that can withstand the pressure when the frit and the capillary column are installed.

Further, the inner peripheral portions of both ends of the main body have threaded screw portions carved respectively. When a screw thread is provided on the inner peripheral portion, it may be appropriate to provide a hole one size larger than the small diameter hole and engrave the screw screw in the large hole.

<Frit> Next, the frit in the internal through hole will be described.

The material of the frit is as follows: 1) having electrical insulation properties; 2) not being corroded by contact with an electrolyte; 3) sufficiently enduring Joule heat generated in a column; 4) being porous as a frit. There is no particular limitation as long as it has properties and can be molded.

Various glasses such as quartz and hard glass,
Various resins such as polyethylene, polypropylene, polystyrene, polyvinyl chloride, polyethylene terephthalate, polytetrafluoroethylene, polyimide, polyamide, polyetheretherketone, phenolic resin, and fluororesin; various ceramics such as alumina, zirconia, titania, and silicon nitride And these may be used singly or in combination of two or more depending on the conditions of use. Of these, resins are preferably used because of ease of molding, and polyetheretherketone is more preferably used.

The frit has a porosity for holding the filler. The porosity as referred to herein may be any as long as the diameter of the pores can hold the filler smaller than the particle diameter of the filler used.
It can be appropriately selected from the range of about 5 μm, and the frit hole may have a hollow hole penetrating the frit in the flow direction of the mobile phase.

The thickness of the frit in the direction of flow of the electrolyte is not particularly limited as long as it does not hinder the spread of the peak of the sample component in the measurement, but is about 0.1 to 1.5 mm.
It is desirable to select from the range. The diameter of the frit may be any size as long as it fits in the internal through-hole, but is preferably the same as or slightly larger than the diameter of the capillary column or capillary tube.

Specific examples of the shape of such a frit include the shapes shown in FIGS. 2, 3, 4 and 5. As a method of using the same, it is preferable to arrange the small-diameter hole of the column connection joint so as to come into contact with the projection in the small-diameter hole from the capillary column side, and then insert the capillary column into the small-diameter hole. In the case of the frit as shown in FIGS. 4 and 5, it is sufficient to insert the small-diameter portion into the small-diameter hole with the small-diameter portion first, and hook the convex portion.

<Screw> Next, a screw having a through-hole inside the column connection joint will be described.

One screw is used for each end of the connection joint body, and the shape and material may be different at both ends.

As the material of the screw, similarly to the material of the column connection joint body described above, it has an electrical insulation property,
There is no particular limitation as long as it can sufficiently withstand Joule heat generated in the column and can be formed as a screw.

For example, polyethylene, polypropylene,
Various resins such as polystyrene, polyvinyl chloride, polyethylene terephthalate, polytetrafluoroethylene, polyimide, polyamide, polyetheretherketone, phenolic resin, fluororesin, alumina, zirconia,
Depending on the structure and shape of the screw, these may be used alone or in combination of two or more depending on the structure and shape of the screw. Of these, resins are preferably used because of ease of molding, and polyetheretherketone is more preferably used.

This screw has a through hole inside, and one of the through holes is for inserting a capillary tube for the sample injection side electrode tank or a capillary tube for the detector side electrode tank. The size can be appropriately selected according to the thickness of the capillary tube to be used, as in the case of the small diameter hole of the column connection joint body described above. The other of the through holes is for inserting a capillary column, and in this case, the size of the hole can be appropriately selected according to the thickness of the capillary column to be used, similarly to the small diameter hole of the column connection joint main body. .

The length of the screw is not particularly limited as long as it has a structure complementary to the cut portion of the column connection joint main body and can fix the capillary tube and the capillary column.

<Ferral> Next, the ferrule constituting the column connection joint will be described.

The ferrule interposed between the column connecting joint body and the screws respectively screwed to both ends thereof is used for each one of the two ends of the column connecting joint body. And the material may be different at both ends.

The material of the ferrule, as with the screw material described above, has electrical insulation properties, can withstand Joule heat generated in the column sufficiently, and can be molded as a ferrule. There is no particular limitation.

For example, polyethylene, polypropylene,
Polystyrene, polyvinyl chloride, polyethylene terephthalate, polytetrafluoroethylene, polyimide, polyamide, polyetheretherketone, phenolic resin, can be appropriately selected from various resins such as a fluororesin, and these can be used as needed. Not only one kind alone, but two or more kinds may be used in combination. Of these, resins are preferably used because of ease of molding, and polyetheretherketone is more preferably used.

The shape of the ferrule is not particularly limited and can be appropriately selected according to the purpose. Further, if necessary, it may be different at both ends of the column connecting joint.

<Capillary tube> Next, the capillary tube used for each electrode tank will be described.

The capillary tube connects the electrode tank used in the present invention and the capillary column, and transfers the electrolytic solution. As its structure and material,
There is no particular limitation as long as it has a thin tube structure and has flexibility so that it can be bent to some extent.

For example, it is possible to use a hollow capillary tube made of various kinds of glass such as made of fused silica and borosilicate glass, and various kinds of resin such as Teflon and polyetheretherketone. In addition, the inner wall is preferably one that does not substantially adsorb the sample component injected or separated by the capillary column. For example, it may be preferable that the inner wall is subjected to an inactivation treatment that does not generate EOF. It is preferable that the capillary tube does not have various liquid coatings as a stationary phase on the inner wall or a solid adsorbent is not fixed, and a so-called empty column can be used. Various capillary columns having phases can also be used.

The length of the capillary tube can be appropriately selected according to the distance between the electrode tanks of the apparatus used and the length of the capillary column, but is preferably about 3 to several tens cm. More preferably, it is about 5 to several tens cm.

The thickness of the capillary tube is not particularly limited as long as it can be connected to the sample-side electrode tank and the detector-side electrode tank, and can be appropriately selected according to the purpose. 0.4 mm, inner diameter 0.02
It can be selected from a range of about 0.1 mm.

The capillary tube connects the sample-side electrode tank to the inlet side of the capillary column via a column connection joint, and the capillary tube connects the outlet side of the capillary column to the detector-side electrode tank via a connection joint. Are required, and at least two tubes are used in one apparatus, but the length and shape of each capillary tube may be different.

<Capillary Column> Next, a capillary column filled with a filler will be described.

The capillary tube used for the capillary column is similar to the above-mentioned capillary tube, and is not particularly limited as long as it has a thin tube structure and has a flexibility that can be bent to some extent.

For example, a hollow capillary tube made of various kinds of glass such as fused silica and borosilicate glass, and various resins such as Teflon and polyetheretherketone can be used. In some cases, it is preferable that the inner wall does not adsorb the sample component injected or separated by the capillary column. The tube preferably has no liquid coating as a stationary phase on the inner wall or has no solid adsorbent fixed thereon, and a so-called empty column can be used.

The length of the capillary column can be appropriately selected according to the distance between the electrode tanks of the apparatus to be used and the length of the capillary tube.
It is about 1 m, more preferably about 5 to several tens cm.

The thickness of the capillary column is not particularly limited as long as it can be connected to the column connection joint, and can be appropriately selected according to the purpose. Preferably, the outer diameter is about 0.25 to 0.4 mm, and the inner diameter is about 0.1 to 0.4 mm. 02-0.1mm
You can choose from a range of degrees.

The type of the filler to be packed in the capillary column is not particularly limited as long as it is a stationary phase that can be used in HPLC. Mold filler, gel permeation filler and the like.

The method of filling the capillary tube with the filler for HPLC is not particularly limited as long as the capillary tube can be filled with the filler, and a dry filling method or a wet filling method can be appropriately selected depending on the purpose. Preferably, the wet filling method may be desirable in consideration of the filling efficiency and the effect on the functional group of the filler.

<Method of Connecting Capillary Tube and Capillary Column> Next, a method of connecting each capillary tube and capillary column will be described.

By inserting the capillary tube and the capillary column into the column connection joint and tightening them with screws screwed to both ends of the column connection joint body, the ferrule interposed therebetween is pressed,
The joint for column connection, the capillary tube and the capillary column can be securely connected and hermetically sealed.

The connection between the capillary tube and the capillary column is carried out through a column connection joint. To prevent a decrease in the separation ability due to the diffusion of the sample in the capillary column, the capillary tube inserted into the column connection joint should be connected to the column as much as possible. It may be preferable to insert up to near the frit inside the joint.

When a column connection joint is used, for example, the first capillary tube is connected to the sample-side electrode tank of the apparatus, and then the capillary tube and the capillary column inlet filled with the filler are connected using the column connection joint. The other side is connected, a second capillary tube is connected to the capillary column outlet side using another connection joint, and this capillary tube is further connected to the detector side device. That is, when connecting a capillary column filled with a filler using a column connection joint, at least two column connection joints are used in one apparatus. Each connection joint may be different in material, shape, and the like. Note that the connection order described above may be appropriately changed according to the work procedure.

<Capillary tube and / or capillary column connected by column connection joint>
Using the various components described above, a capillary tube and / or a capillary column can be obtained by connecting the column connecting joint of the present invention via a frit. This column can be incorporated into the device without reducing the separation capacity due to CEC. In addition, this column can be used by connecting the columns as necessary and combining capillary columns filled with various fillers. Furthermore, this column can be stored for a long period of time under conditions where there is no deterioration of the packing material, and can be used by incorporating it into the device when necessary.

<CEC Device> The column connected by the column connection joint described above can be used as a CEC device by combining the components shown in FIG.

The CEC apparatus comprises a capillary column 8 having column connection joints 7a and 7b according to the present invention.
Capillary tube 6 for sample injection side electrode tank, capillary tube 9 for detector side electrode tank, high voltage power supply 1 and power cable 2, electrode 3, sample injection side electrode tank 4, detector side electrode tank 5, and detection cell 11 It has a detector 10 and a data processing device 12.

1) Capillary tube The capillary tube 9 for the electrode tank on the detector side is a detector 10
The sample-injection-side electrode tank capillary tube 6 connects the capillary column 8 and the sample-injection-side electrode tank 4 with each other.
Any of these can be known.

2) Sample Injection-Side Electrode Tank and Detector-Side Electrode Tank There is no particular limitation as long as the object of the present invention can be achieved.
Known ones can be used.

3) Electrodes Known electrodes can be used which are installed in the sample-injection-side electrode tank and the detector-side electrode tank.

4) High Voltage Power Supply The high voltage power supply used in the present invention has a voltage of about 3
The device is not particularly limited as long as the device has an output up to 0 kV and a current in the range of 100 to 300 μA.
It can be appropriately selected according to the purpose, and preferably, the output electrode can be inverted according to the measurement sample. In addition, as the power supply cable, any known high-voltage power supply can be used as long as the high-voltage power supply used properly operates and electricity can be transmitted to the electrodes 3.

5) Detector The detector 10 used in the present invention is not particularly limited as long as it can detect a sample component. An ultraviolet-visible detector, a multi-wavelength detector, a fluorescence detector, an electric detector Examples include a conductivity detector and a differential refraction detector. Further, the detection cell 11 may be a cell corresponding to the detector, and a known cell can be used.

6) Data Processing Apparatus As the data processing apparatus 12, any known apparatus can be used as long as it can record and process the data obtained from the detector. Any processing device that can achieve the object of the present invention, such as a device that has an output function and can be processed by a computer, can be adopted.

7) Sample Injection Method The sample injection method used in the present invention is not particularly limited as long as a sample component can be injected into a capillary tube, and includes a drop method, an electrophoresis method, a pressurization method, and the like.
A suction method and the like can be mentioned, and it can be appropriately selected according to the purpose.

[0063]

Embodiments of the present invention will be described below with reference to the drawings, but the present invention is not limited to only these embodiments.

Example 1 FIG. 1 is a block diagram of a capillary electrochromatography apparatus of the present invention. In FIG. 1, common components and mutual relationships are denoted by common reference numerals, and description thereof is omitted.

Here, in FIG. 1, 1 is a high-voltage power supply, and 2 and 3 are voltage cables and electrodes connected to the high-voltage power supply. Reference numerals 4 and 5 denote a sample injection-side electrode tank and a detector-side electrode tank. Reference numeral 6 denotes a capillary tube for a sample injection side electrode tank, 7a and 7b are connection joints having a frit therein, 8 is a capillary column filled with a filler, and 9 is a capillary tube for a detector side electrode tank. is there. 10 and 11 are a UV detector and a detection cell, and 12 is a data processing device.

As shown in FIG. 1, in the capillary electrochromatography apparatus of the present invention, the connection joint 7 having a frit inside needs to be connected to both sides of the capillary column 8 filled with the packing material.
A minimum of 2 per unit is required.

Example 2 FIG. 6 is a schematic diagram of an axial cross section of a column connection joint for connecting a capillary tube and a capillary column used in the capillary electrochromatography apparatus of the present invention. In FIG. 6, common parts and mutual relations are denoted by common reference numerals and description thereof is omitted, and the same applies to FIGS. 7, 8 and 9 described later.

In FIG. 6, reference numeral 13 denotes a connecting joint main body, which is connected to the capillary tube 2 on one side.
4 and a small hole 15 for connecting a capillary column 25 to the other. These small holes are connected to each other, and An internal through hole 16 is formed on the inside, and screwing screws 17 and 1 are formed on inner peripheral portions at both ends.
8 is engraved. Reference numeral 19 denotes a frit for holding the filler, and FIG. 6 uses a frit 19 having the shape shown in FIGS. 2 and 3; 20 and 21 are screws having through holes, respectively; And 23 are ferrules interposed between the column connection joint main body and the screws, and 24 is an electrode tank side (including a sample electrode tank).
Or a capillary tube on the electrode tank side for the detector,
25 is a capillary column, and 26 is a packing material for HPLC.

As shown in FIG. 6, the column connecting joint, the capillary tube 24 and the capillary column 25
Are connected, and the screws 20 and 21 having through holes are tightened, respectively, so that the ferrules 22 and 23 are deformed and can be tightly fixed to the column connection joint body 13 respectively.

Embodiment 3 In FIG. 7, the inner diameter of the projection 27 in the column connection joint body 13 of FIG. 6 of the embodiment 2 is increased, and the capillary tube 24 is made to penetrate the projection 27 in FIGS. Is in direct contact with the frit 19 having the shape shown in FIG.
In FIG. 7, the capillary tube 24 and the capillary column 25 can be interchanged.

Example 4 FIG. 8 shows an example in which the capillary tube 24 is replaced with a capillary column 25 in FIG. 7 of Example 3 and two capillary columns 25 are used.

Embodiment 5 In FIG. 9, a frit having the shape shown in FIGS. 4 and 5 is used in place of the frit in FIG.
9 in direct contact.

Example 6 After adding an electrolytic solution to benzyl alcohol, it was completely dissolved with a shaker to obtain a measurement sample. CEC measurement conditions include a high voltage power supply (890-C, manufactured by JASCO Corporation).
E), TSKgel G3000SWX as filler
L (manufactured by Tosoh), a capillary column (inner diameter 0.1 mm, length 15 cm), a fused silica tube (inner diameter 0.1 mm, length 5 cm) as a capillary tube for the sample injection side electrode tank, and a detector side electrode tank A fused silica tube (inner diameter 0.1 mm, length 20 cm) was used as a capillary tube, and polyether ether ketone was used as a material for a connection joint having a frit inside. An ultraviolet-visible detector (870-CE, manufactured by JASCO Corporation) was used as the detector, and the measurement wavelength was 200.
nm was used. As the electrolyte, 50 mM disodium hydrogen phosphate (pH 7.0) was used. As a sample injection method, an applied voltage of 5 was applied by using a drop method (5 cm, 10 sec).
Electrophoresis was performed at kV and an applied current of 50 μA. The chromatogram obtained from the result is shown in FIG.

Example 7 Measurement was performed by CEC under the same measurement conditions as in Example 6. As a measurement sample, a mixture of bovine serum albumin, ovalbumin (ovalbumin), myoglobin and p-aminobenzoic acid was used. First, after the electrolyte solution was added to the mixture, it was completely dissolved with a shaker to obtain a measurement sample. The detector used was an ultraviolet-visible detector 870-CE, and the measurement wavelength was 200
nm. The electrolyte used was 20 mM sodium dihydrogen phosphate (pH 2.4). The sample was injected using a manual pressurization method, with an applied voltage of 5 kV and an applied current of 36μ.
A electrophoresis was performed. The resulting chromatogram is shown in FIG.

[0075]

As described above, the conventional capillary column structure is divided into a capillary tube for an electrode tank on the sample injection side, a capillary column filled with a packing material, and a capillary tube for an electrode tank on the detector side, each of which is internally provided. By connecting with a connection joint having a frit, 1) the operation of preparing a frit and a detection cell for each capillary column is omitted, and the packing and preparation method is simplified. 2) The exchange operation of the capillary column is facilitated. 3) CHPLC However, it can be applied as an analytical column, and is of great practicality.

[Brief description of the drawings]

FIG. 1 is an example of a configuration diagram of a capillary electrophoresis apparatus of the present invention.

FIG. 2 is a schematic diagram example of a frit used in a column connection joint of the present invention as viewed from a cross-sectional direction.

FIG. 3 is a schematic diagram example of a frit used in a column connection joint of the present invention as viewed from a slope direction.

FIG. 4 is a schematic diagram example of a frit used in a column connection joint of the present invention as viewed from a cross-sectional direction.

FIG. 5 is a schematic diagram showing an example of a frit used in the column connection joint of the present invention as viewed from a slope direction.

FIG. 6 is an example of a schematic diagram of a cross section in the axial direction of a connection between a capillary column and a capillary tube using the column connection joint of the present invention.

FIG. 7 is an example of a schematic diagram of an axial cross section of a connection between a capillary column and a capillary tube using the column connection joint of the present invention.

FIG. 8 is a schematic diagram illustrating an example of a cross section in the axial direction of a capillary column connected using the column connection joint of the present invention.

FIG. 9 is a schematic diagram of an axial cross section of a capillary column and a capillary tube connected using the column connection joint of the present invention.

FIG. 10 is a chromatogram obtained in Example 6,
The X axis (horizontal axis) is the retention time in CEC measurement (unit is
Minutes), and the Y axis (vertical axis) indicates the intensity of the obtained peak (arbitrary unit).

FIG. 11 is a chromatogram obtained in Example 7,
The X axis (horizontal axis) is the retention time in CEC measurement (unit is
Minutes), and the Y axis (vertical axis) indicates the intensity of the obtained peak (arbitrary unit).

[Explanation of symbols]

The numbers in the figure are common, and the meanings of the numbers are shown below. 1: High voltage power supply 2: High voltage cable 3: Electrode 4: Sample injection side electrode tank 5: Detector side electrode tank 6: Capillary tube for sample injection side electrode tank 7a, b: Connection having frit inside Joint 8: Capillary column filled with packing material 9: Capillary tube for detector-side electrode tank 10: Ultraviolet-visible detector 11: Detection cell 12: Data processing device 13: Column connection joint body 14: Connecting capillary tube Small hole 15: Small hole for connecting a capillary column 16: Internal through hole 17, 18: Screw screw 19: Frit 20, 21: Screw having a through hole 22, 23: Ferrule 24: Capillary tube 25: Capillary column 26: Packing material 27: Convex part in column connection joint body 28: Benzil Peak of alcohol 29: bovine serum albumin peak 30: Peak myoglobin 31: peak of ovalbumin 32: Peak of p- acid

Claims (8)

[Claims] 1. One has a small diameter hole for connecting a capillary column filled with a packing material, and the other has a small diameter hole for connecting a capillary tube, and these small diameter holes are mutually connected. A column connection joint body that is connected to form an internal through hole, a frit for holding a filler is installed in the internal through hole, and a screw screw is engraved on each of the inner peripheral portions at both ends. A screw having a through-hole that is screwed to each end of the column connection joint body, and ferrules interposed between the column connection joint body and the screw, respectively. Column connection joint. 2. The column connection joint according to claim 1, further comprising a projection for holding a frit in the internal through hole. 3. A column connection joint for connecting a capillary column of capillary electrochromatography, wherein the material of the column connection joint has electrical insulation and is made of glass, resin and ceramics. The joint for column connection according to claim 1 or 2, wherein the joint is a single member selected from the group consisting of a combination of two or more members. 4. A capillary column, wherein the column connection joint according to claim 1 is connected to a capillary column. 5. The capillary column according to claim 4, wherein the packing is a stationary phase for high performance liquid chromatography. 6. A capillary for migrating a sample, a sample injection-side electrode tank and a detector-side electrode tank provided on both sides of the capillary, and connected to the electrodes provided in these electrode tanks via cables. In a capillary electrochromatography apparatus including a high-voltage power supply for causing a sample to migrate in a capillary and a detection unit for detecting the sample provided in the middle of the capillary, the capillary is filled with a filler. Capillary column, and divided into a capillary tube for the sample injection side electrode tank and a capillary tube for the detector side electrode tank on both sides of the capillary column, the capillary column and the capillary tube according to any one of claims 1 to 3 It is characterized by being connected via a column connection joint. Capillary electrochromatography device to. 7. A capillary electric device according to claim 6, wherein the capillary column has at least two capillary columns, and the capillary columns are connected to each other via the connection joint according to any one of claims 1 to 3. Chromatography equipment. 8. The capillary electrochromatography apparatus according to claim 6, wherein the packing material packed in the capillary column is a stationary phase for high performance liquid chromatography.